Nucleic acids encoding the GPCR, RUP3, and methods of use thereof

• US 7,893,235 B2
• Filed: 08/20/2007
• Issued: 02/22/2011
• Est. Priority Date: 11/20/1998
• Status: Expired due to Fees

• Current Assignee: Arena Pharmaceuticals Incorporated (Pfizer Inc.)
• Original Assignee: Arena Pharmaceuticals Incorporated (Pfizer Inc.)
• Inventors: Liaw, Chen W., Chen, Ruoping, Lin, I-Lin, Dang, Huong T.
• Primary Examiner(s): LANDSMAN, ROBERT S
• Application Number:
  US11/894,692
• Publication Number:
  US 20080199889A1
• Time in Patent Office: 1,282 Days
• Field of Search: None
• US Class Current: 536/23.5
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 14/723 G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor
• CPC Class Codes: G01N 2333/726 G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
• CPC Class Codes: G01N 2500/02 Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)
• CPC Class Codes: G01N 33/74 involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors

Abstract

The invention disclosed in this patent document relates to transmembrane receptors, more particularly to endogenous, human orphan G protein-coupled receptors.

Claims

1. An isolated nucleic acid which encodes a G protein-coupled receptor, wherein said nucleic acid comprises SEQ ID NO:

7.

12. A vector comprising the isolated nucleic acid according to any one of claim 1, 2, 3, 4-7, 8-10 or 11.

13. A plasmid comprising the isolated nucleic acid according to any one of claim 1, 2, 3, 4-7, 8-10 or 11.

14. An isolated host cell comprising the vector according to claim 12 or the plasmid according to claim 13.

15. The isolated host cell according to claim 14, wherein said isolated host cell is a eukaryotic host cell.

16. The isolated host cell according to claim 15, wherein said eukaryotic host cell is a mammalian host cell.

17. The isolated host cell according to claim 16, wherein said mammalian host cell is selected from the group consisting of:

293, 293T and COS-7 cells.

18. A method of producing a G protein-coupled receptor encoded by an isolated nucleic acid according to any one of claim 1, 2, 3, 4-7, 8-10 or 11, said method comprising the steps of:

a) transfecting a host cell with a vector encoding a G protein-coupled receptor encoded by an isolated nucleic acid according to any one of claim 1, 2, or 3, 4-7, 8-10 or 11; and

b) culturing the transfected host cell under conditions sufficient to express the G protein-coupled receptor from the vector.

19. A method for identifying a compound for inhibiting or stimulating a G protein-coupled receptor encoded by an isolated nucleic acid according to any one of claim 1, 2, 3, 4-7, 8-10 or 11, said method comprising the steps of:

a) contacting one or more candidate compounds with a host cell or membrane thereof, wherein said host cell or membrane comprises a G protein-coupled receptor encoded by an isolated nucleic acid according to any one of claim 1, 2, 3, 4-7, 8-10 or 11; and

b) measuring the ability of the candidate compound or compounds to inhibit or stimulate said G protein-coupled receptor.

20. A method for identifying a compound capable of binding to a G protein-coupled receptor encoded by an isolated nucleic acid according to any one of claim 1, 2, or 3, 4-7, 8-10 or 11, said method comprising the steps of:

a) contacting one or more candidate compounds with a host cell or membrane thereof, wherein said host cell or membrane comprises a G protein-coupled receptor encoded by an isolated nucleic acid according to any one of claim 1, 2, 3, 4-7, 8-10 or 11; and

b) measuring the ability of the candidate compound or compounds to bind to said G protein coupled receptor.

2. An isolated nucleic acid which encodes a G protein-coupled receptor, wherein said G protein-coupled receptor comprises the amino acid sequence of SEQ ID NO:

8.

3. An isolated nucleic acid which encodes a G protein coupled receptor, wherein said G protein-coupled receptor comprises the amino acid sequence of a mutant of SEQ ID NO:

8 comprising a substitution of lysine for leucine at amino acid position 224 of SEQ ID NO;

8.

4. An isolated nucleic acid which encodes a G protein-coupled receptor with at least 95% amino acid sequence identity to SEQ ID NO:

8, wherein said G protein-coupled receptor is capable of modulating insulin or glucagon levels.

5. The nucleic acid according to claim 4, wherein said nucleic acid encodes a G protein-coupled receptor with at least 98% amino acid sequence identity to SEQ ID NO:

8.

6. The nucleic acid according to claim 4 or 5, wherein said nucleic acid encodes a constitutively activated G protein-coupled receptor.

7. The nucleic acid according to claim 4 or 5, wherein said nucleic acid encodes a human G protein-coupled receptor.

8. An isolated nucleic acid which encodes a G protein-coupled receptor with at least 95% amino acid sequence identity to SEQ ID NO:

8, wherein said G protein-coupled receptor is a RUP3 protein.

9. The nucleic acid according to claim 8, wherein said nucleic acid encodes a G protein-coupled receptor with at least 98% amino acid sequence identity to SEQ ID NO:

8.

10. The nucleic acid according to claim 8 or 9, wherein said nucleic acid encodes a constitutively activated G protein-coupled receptor.

11. The nucleic acid according to claim 8 or 9, wherein said nucleic acid encodes a human G protein-coupled receptor.